Publications

Here is a selection of publications where different laminin isoforms are being used to create more authentic cell culture systems

  • Area of interest

  • Regulation of the Immune System by Laminins

    Simon T., Bromberg J. Trends Immunol, 2017

    This review summarizes the structure of laminins, the modulation of their expression, and their interactions with the immune system. In addition, the role of laminins in autoimmune diseases and transplantation are discussed.

  • A Chemically Defined Hydrogel for Human Liver Organoid Culture

    Ye S., Boeter J.W.B., Mihajlovic M., van Steenbeek F.G, van Wolferen M.E., Oosterhoff  L.A., Marsee A., Caiazzo M., van der Laan L.J.W., Penning L.C., Vermonden T., Spee B., and Schneeberger K. Adv. Funct. Mater. 2020

    Here, a novel hydrogel-based on polyisocyanopeptides (PIC) and Biolaminin-111 is described for human liver organoid cultures. PIC is a synthetic polymer that can form a hydrogel with thermosensitive properties, making it easy to handle and very attractive for clinical applications. PIC hydrogel alone was not sufficient to support organoid growth. The addition of a laminin-entactin complex (LEC) to the plain PIC gel, resulted in efficient organoid formation and proliferation that seemed comparable to the Matrigel controls, with lower stiffnesses most favorable for organoid proliferation. The stem cell phenotype and proliferation and differentiation capacity of the organoids could be maintained in PIC-LEC over several passages, enabling their seemingly unlimited expansion and subsequent maturation. Moreover, organoids can be efficiently differentiated toward a hepatocyte-like phenotype with key liver functions. Importantly, they also show that the LEC in the PIC-LEC gels could be replaced by Biolaminin-111, resulting in a completely synthetic hydrogel for the expansion of human liver organoids.

  • The ability of inner-cell-mass cells to self-renew as embryonic stem cells is acquired following epiblast specification

    Boroviak T., Loos R., Bertone P., Smith A. and Nichols J.Nature Cell Biology, 2014

    The authors show that mouse ICM cells from early blastocysts can progress to ERK independence if provided with a specific laminin substrate. We show by RNA sequencing that Laminin-511 is expressed in the early ICM, as is B1-integrin, which mediates laminin binding in ESCs. No expression of Laminin-521 in mouse ICM. Single ESC derivation in 2i-LIF on a fibronectin-LN511 mix and clonal ESC lines expansion on LN511.

  • A Balance between Secreted Inhibitors and Edge Sensing Controls Gastruloid Self-Organization

    Etoc F., Metzger J., Ruzo A., Kirst C., Yoney A., Ozair Z.M, Brivanlou A.H., Siggia E.D. Developmental cell, 2016

    Used colonies of hESC grown on the micropatterned substrate and differentiated with BMP4 to model patterning events in the human embryo. Coated Costar Transwells with LN521.

  • CCR2+CCR5+ T Cells Producing Matrix Metalloproteinase-9 and Osteopontin in the Pathogenesis of Multiple Sclerosis

    Sato W., Tomita A., Ichikawa D., Lin Y., Kishida H., Miyake S., Ogawa M., Okamoto T., Murata M., Kuroiwa Y., Aranami T., Yamamura T.Journal of Immunology, 2012

    To recapitulate the glia limitans layered with parenchymal basal lamina experimentally, we coated the upper sides of Transwell membrane inserts. The upper sides of Transwell membrane inserts (8 mm; Corning) were coated with 10 mg/ml laminin-1 (Sigma) or 20 mg/ml laminin-121. After aspirating the laminin solutions, the membrane inserts were turned upside down, and normal human astrocytes (NHA) were seeded on the lower sides of the membrane inserts. T cells were stimulated, harvested, suspended in the fresh medium, and seeded onto the upper chambers. After 8 h, the cell suspension was collected from the lower chambers after careful pipetting, and absolute numbers of migrated cells were calculated. The T-cell migration across the NHA layered with laminin-111 or -121 was less efficient compared with the migration across the untreated membrane or the membrane treated with laminin alone, thus, this model would exhibit barrier functions against the penetration of activated T cells.

  • Early embryonic-like cells are induced by downregulating replication-dependent chromatin assembly

    Ishiuchi T., Enriquez-Gasca R., Mizutani E., Bošković A., Ziegler-Birling C., Rodriguez-Terrones D., Wakayama T., Vaquerizas J.M., Torres-Padilla M-E.Nature structural & molecular biology, 2015

    mES cells cultured on coverslips coated with laminin-511for an RNA-FISH assay.

  • Dynamic Heterogeneity and DNA Methylation in Embryonic Stem Cells

    Singer Z.S., Yong J., Tischler J., Hackett J.A., Altinok A., Surani M.A., Cai L., and Elowitz M.B.Cell Press, 2014

    Analyzis of gene expression in individual ESCs using single-molecule RNA-FISH and quantitative time-lapse movies. Fluorescence Time-Lapse Microscopy and Data analysis on reporter cells mixed with unlabeled parental cells at 1:9 ratio and plated at a total density of 20,000 cells/cm2 on glass-bottom plates (MatTek) coated with human laminin-511 >12 hr before imaging.

  • Control of ground-state pluripotency by allelic regulation of Nanog

    Miyanari Y., Torres-Padilla M.E.Nature Letter, 2012

    Laminin-511 used to coat glass-bottomed dishes.

  • Live visualization of chromatin dynamics with fluorescent TALEs

    Miyanari Y., Ziegler-Birling C., Torres-Padilla M.E.Nature structural & molecular biology, 2013

    The authors of these two Nature publications show that laminin can be coated directly on glass, which many other substrates and proteins can not. This enables the growth of pluripotent stem cells as monolayers even on glass, which is especially suitable for live imaging.

  • Improved Human Pluripotent Stem Cell Attachment and Spreading on Xeno-Free Laminin-521-Coated Microcarriers Results in Efficient Growth in Agitated Cultures

    Alan Tin-Lun Lam, Jian Li, Allen Kuan-Liang Chen, William R Birch, Shaul Reuveny, Steve Kah-Weng Oh. Biores Open Access, 2015

    The authors used different bioreactors up to 2.5 L in scale, and successfully cultured hMSCs using the minimum agitator speed required for complete microcarrier suspension. In addition, we also reported a scaleable protocol for the detachment from microcarriers in spinner flasks where they use a short period of intense agitation in the presence of enzymes such that the cells are detached without being damaged. Here, the same approach has been effective for 15 mL ambrTMbioreactors, 100 mL spinner flasks and 250 mL Dasgip bioreactors.  Two types of microcarrier were used, with (laminin-521) and without surface coatings, four different enzymes and three different growth media (with and without serum), a total of 22 different combinations. In all cases after detachment, the cells were shown to retain their desired quality attributes and were able to proliferate. Qasim Rafiq presented this data at BioLamina symposium 2015. It was found that human recombinant laminin LN-521 and recombinant fibronectin were the optimal ECM proteins for microcarrier coating. A serum-free expansion, harvest and xeno, and DMSO-free cryopreservation process, using LN-521 coated microcarriers and FREEZEstemTM cryopreservation medium was then developed which demonstrated > 5 fold increase in hMSC yield.